Online citations, reference lists, and bibliographies.
← Back to Search

Site-Directed Pegylation Of Recombinant Interleukin-2 At Its Glycosylation Site

R. Goodson, N. Katre
Published 1990 · Chemistry, Medicine

Save to my Library
Download PDF
Analyze on Scholarcy
Share
We have modified recombinant interleukin-2 (rIL-2) to facilitate site-directed covalent attachment of monomethoxy polyethylene glycol (PEG). The site chosen for modification and subsequent covalent attachment with PEG (PEGylation) was the single glycosylation position found in the native interleukin-2 (IL-2). The mutant protein was expressed in E. coli, purified, and PEGylated with a PEG-maleimide reagent to obtain PEG-cys3-rIL-2. The PEG-cys3-rIL-2 had full bioactivity relative to the unmodified molecule and had an increase in hydrodynamic size sufficient to increase its systemic exposure by ∼4 fold. This method has general applicability for modifying any therapeutic protein at a specific site and thereby alter its potency. In particular, it can be used to attach PEG to prokaryotically expressed recombinant proteins at their glycosylation sites.
This paper references
10.1016/0003-2697(83)90131-8
A new procedure for the synthesis of polyethylene glycol-protein adducts; effects on function, receptor recognition, and clearance of superoxide dismutase, lactoferrin, and alpha 2-macroglobulin.
C. Beauchamp (1983)
10.1126/SCIENCE.3131876
Interleukin-2: inception, impact, and implications.
K. Smith (1988)
10.1016/S0075-7535(08)70259-6
Chemical modification of proteins: Selected methods and analytical procedures
A. Glazer (1975)
10.1093/NAR/13.12.4431
Improved oligonucleotide site-directed mutagenesis using M13 vectors.
P. Carter (1985)
Cancer therapy with chemically modified enzymes. I. Antitumor properties of polyethylene glycol-asparaginase conjugates.
A. Abuchowski (1984)
10.1073/PNAS.84.6.1487
Chemical modification of recombinant interleukin 2 by polyethylene glycol increases its potency in the murine Meth A sarcoma model.
N. Katre (1987)
10.1021/JO01314A032
New, easily removable poly(ethylene glycol) supports for the liquid-phase method of peptide synthesis
V. Pillai (1980)
10.1093/NAR/12.24.9441
The gapped duplex DNA approach to oligonucleotide-directed mutation construction.
W. Kramer (1984)
10.1016/0003-2697(87)90524-0
A water-soluble, monitorable peptide and protein crosslinking agent.
L. Aldwin (1987)
10.1016/0022-1759(86)90183-3
An improved colorimetric assay for interleukin 2.
H. Tada (1986)
10.1016/s0021-9258(18)68146-3
Relationship of effective molecular size to systemic clearance in rats of recombinant interleukin-2 chemically modified with water-soluble polymers.
M. J. Knauf (1988)
10.1038/NBT1186-1006
High–Level Expression of Diphtheria Toxin Peptides in Escherichia coli
L. Greenfield (1986)
10.1126/SCIENCE.6367046
Biological activity of recombinant human interleukin-2 produced in Escherichia coli.
S. Rosenberg (1984)
Schedule dependency of the antitumor activity and toxicity of polyethylene glycol-modified interleukin 2 in murine tumor models.
R. J. Zimmerman (1989)
10.1016/S0076-6879(85)16040-4
Human interleukin 2.
R. Robb (1985)
Recent advances in the preclinical and clinical immunopharmacology of interleukin-2: emphasis on IL-2 as an immunorestorative agent.
Hadden Jw (1988)
10.1073/PNAS.81.20.6486
Amino acid sequence and post-translational modification of human interleukin 2.
R. Robb (1984)
10.1126/SCIENCE.6427925
Site-specific mutagenesis of the human interleukin-2 gene: structure-function analysis of the cysteine residues.
A. Wang (1984)
10.1016/S0022-2836(83)80284-8
Studies on transformation of Escherichia coli with plasmids.
D. Hanahan (1983)
Immunogenicity of recombinant IL-2 modified by covalent attachment of polyethylene glycol.
N. Katre (1990)
10.1016/0076-6879(83)00074-9
Oligonucleotide-directed mutagenesis of DNA fragments cloned into M13 vectors.
Mark J. Zoller (1983)
10.1038/NBT0488-417
Stable Expression of Aspercillus Awamori Glucoamylase in Distiller's Yeast
G. Cole (1988)



This paper is referenced by
10.3109/10837450.2010.513990
Product development issues for PEGylated proteins
R. W. Payne (2011)
10.1016/j.exphem.2008.08.003
Erythropoietins: a common mechanism of action.
S. Elliott (2008)
10.1016/0021-9673(92)85467-8
Characterization of the heterogeneity of polyethylene glycol-modified superoxide dismutase by chromatographic and electrophoretic techniques.
J. Snider (1992)
10.1016/J.ADDR.2007.06.015
Site-specific modification and PEGylation of pharmaceutical proteins mediated by transglutaminase.
A. Fontana (2008)
10.1016/j.maturitas.2011.12.014
Nanomedicine for the prevention, treatment and imaging of atherosclerosis.
C. Psarros (2012)
10.1016/B978-0-444-53349-4.00101-1
Oligomeric Poly(ethylene oxide)s. Functionalized Poly(ethylene glycol)s. PEGylation
I. Dimitrov (2012)
10.1016/J.ADDR.2007.02.001
Releasable PEGylation of proteins with customized linkers.
D. Filpula (2008)
UNIVERSIT DE BORDEAUX U.F.R DES SCIENCES PHARMACEUTIQUES
Par Marie-Anne Guitou (2014)
10.1016/j.bbagen.2013.03.031
Quantification of thiols and disulfides.
J. R. Winther (2014)
10.1016/J.PROGPOLYMSCI.2006.03.001
Michael addition reactions in macromolecular design for emerging technologies
B. D. Mather (2006)
Developing Novel Electrospray Ionization Mass Spectrometry (esi ms) Techniques to Study Higher Order Structure and Interaction of Biopolymers
Agya K. Frimpong (2009)
10.1016/S0040-4039(00)78260-8
Preparation of a new PEGylation reagent for sulfhydryl-containing polypeptide
W. Tang (1994)
10.1201/9781420008357.CH24
PEGylation of Biological Macromolecules
L. Bass (2007)
10.14288/1.0068503
Prevention of respiratory syncytial virus infection via methoxypoly(ethylene glycol)-modification of the virus or its host cell
T. C. Sutton (2009)
10.1007/3-7643-7543-4_15
New molecules and formulations of recombinant human erythropoietin
S. Elliott (2003)
Inhibiteurs sélectifs et puissants de nav1.7
J. Murray (2014)
10.1504/IJNT.2013.058114
Gold nanoparticle-based platforms as cancer-targeted molecules delivery systems
Cheng-Cheung Chen (2013)
10.1007/BF02786956
Reversible modification of thiol-containing polypeptides with poly(ethylene glycol) through formation of mixed disulfide bonds
Tony Musu (1996)
10.1007/s12010-014-1207-5
Formulation and Characterisation of Antibody-Conjugated Soy Protein Nanoparticles—Implications for Neutralisation of Snake Venom with Improved Efficiency
Kadali Renu (2014)
10.22377/AJP.V11I01.1090
Effect of Polyethylene Glycol Chain Length on PEGylation of Dendrimers
H. Khambete (2017)
10.1080/10826068.2015.1135463
Modification with polysialic acid–PEG copolymer as a new method for improving the therapeutic efficacy of proteins
Jianrong Wu (2016)
10.1016/J.YMTHE.2004.02.014
Tumor efficacy and biodistribution of linear polyethylenimine-cholesterol/DNA complexes.
D. Furgeson (2004)
10.1016/S1461-5347(98)00086-8
Polyethylene glycol-conjugated pharmaceutical proteins
P. Bailon (1998)
10.1074/jbc.RA118.004551
Dynamic disulfide exchange in a crystallin protein in the human eye lens promotes cataract-associated aggregation
E. Serebryany (2018)
10.1517/17425240802650568
PEG-modified biopharmaceuticals
P. Bailon (2009)
10.1016/S0169-409X(02)00024-8
Enzymatic procedure for site-specific pegylation of proteins.
H. Sato (2002)
Site-Specific Protein PEGylation: Application to Cysteine Analogs of Recombinant Human Granulocyte Colony-Stimulating Factor.
Mary S. Rosendahl (2005)
Bioresponsive polymer therapeutics containing coiled-coil motifs
Samuel P E Deacon (2009)
10.1074/JBC.271.36.21969
Long-acting Growth Hormones Produced by Conjugation with Polyethylene Glycol*
R. Clark (1996)
10.1016/S0169-409X(02)00023-6
Mono-N-terminal poly(ethylene glycol)-protein conjugates.
O. Kinstler (2002)
10.1016/j.jconrel.2018.01.018
Bioorthogonal strategies for site‐directed decoration of biomaterials with therapeutic proteins
Alexandra Braun (2018)
10.1021/BC049713N
A Long-Acting, Highly Potent Interferon α-2 Conjugate Created Using Site-Specific PEGylation
Mary S. Rosendahl (2005)
See more
Semantic Scholar Logo Some data provided by SemanticScholar